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Trypanosomes Lacking Trypanothione Reductase Are Avirulent and Show Increased Sensitivity to Oxidative Stress by S. Krieger; W. Schwarz; M. R. Ariyanayagam; A. H. Fairlamb; R. L. Krauth-Siegel; C. Clayton is a Biochemistry, Genetics and Molecular Biology article available to read on EtoBox.

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In Kinetoplastida, trypanothione and trypanothione reductase (TRYR) provide an intracellular reducing environment, substituting for the glutathione–glutathione reductase system found in most other organisms. To investigate the physiological role of TRYR in __Trypanosoma brucei__, we generated cells containing just one trypanothione reductase gene, __TRYR__, which was under the control of a tetracycline‐inducible promoter. This enabled us to regulate TRYR activity in the cells from less than 1% to 400% of wild‐type levels by adjusting the concentration of added tetracycline. In normal growth medium (which contains reducing agents), trypanosomes containing less than 10% of wild‐type enzyme activity were unable to grow, although the levels of reduced trypanothione and total thiols remained constant. In media lacking reducing agents, hypersensitivity towards hydrogen peroxide (EC~50~ = 3.5 μM) was observed compared with the wild type (EC~50~ = 223 μM). The depletion of TRYR had no effect on susceptibility to melarsen oxide. The infectivity and virulence of the parasites in mice was dependent upon tetracycline‐regulated TRYR activity: if the trypanosomes were injected into mice in the a

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Author

S. Krieger; W. Schwarz; M. R. Ariyanayagam; A. H. Fairlamb; R. L. Krauth-Siegel; C. Clayton

Publisher

John Wiley and Sons; Wiley (Blackwell Publishing); Blackwell Publishing Inc.; Wiley (ISSN 0950-382X)

Published

2002

Language

EN

Field

Biochemistry, Genetics and Molecular Biology (Life Sciences)